Identification and Characterization of Berberine in Tinospora cordifolia by Liquid Chromatography Quadrupole Time of Flight Mass Spectrometry (LC MS/MS Q-tof) and Evaluation of its anti Inflammatory Potential

Abstract

Introduction: Tinospora cordifolia (Willd.) Miers ex Hook F and Thomas commonly called as gudduchi or amrita is a widely used plant in traditional medicinal system of Ayurveda. A UPLC MS/MS Q-tof method for the identification and characterization of berberine in Tinospora cordifolia (Willd.) Miers. ex HooK.F. and Thomas. and to evaluate the anti inflammatory potential of bioactive fraction. Materials and Methods: The presence of berberine in Tinospora cordifolia was determined by HPLC and was subsequently isolated by HPTLC. The anti inflammatory property of the fraction containing berberine was demonstrated to have an inhibitory activity on 5 lipoxygenase, an enzyme involved in inflammatory pathway and its IC 50 value was obtained. The binding interactions between berberine and 5-LOX were demonstrated by docking studies. Result: The presence of berberine in Tinospora cordifolia methanolic extract was identified by HPLC and HPTLC analysis and confirmed by UPLC MS/ MS Q-tof. The fraction containing berberine inhibited 5-LOX with an IC 50 of 0.041± 0.0003µg/mL as compared to that of NDGA (positive control) which showed an IC 50 of 2.75 ± 0.05 µg/mL. Molecular docking of berberine with 5-LOX showed a binding energy of-8.942 ± 0.039665 kcal/mol and Ki of 273.16 ± 3.026 nM as compared to the NDGA which has a binding energy of-7.186 ± 0.170503 kcal/mol and Ki 5.604± 1.618 µM. Conclusion: Tinospora cordifolia can be used as a source of berberine and possible anti inflammatory activity of Tinospora cordifolia may be attributed to the presence of berberine.

Identification and characterization of bioactive compound berberine in the Berberis vulgaris root extract using HR-LC-MS analysis

“…techniques have been developed for the qualitative and quantitative estimation of berberine in Tinospora cordifolia, various formulation, rat and rabbit plasma. [17][18][19][20][21] But there is no report on the quantitative screening of berberine content in the root extract of B. vulgaris using HR-LC/MS analysis. It has been observed that the vacuum evaporated extract of B. vulgaris gave yellowish dark brown extract which was subjected to HR-LC/MS analysis for proper qualitative as well as quantitative identification of berberine.…”

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“…High-Performance Thin Layer Chromatography: HPTLC fingerprinting analysis of Varanadi kashayam fractions was done according to the protocol described in Mohind et al, with modifications 24 . CAMAG HPTLC system (Switzerland) with a Linomat 5 sample applicator was used for the analysis.…”

Identification of the natural product berberine as an antiviral drug

“…It has also been frequently used for the adjuvant treatment of type 2 diabetes mellitus, ischemic myocardium, hyperlipidemia and hypertension in China (Dong et al 2013 ). There is some evidence that berberine has anti-inflammation (Mohan et al 2017 ), anti-aging properties (Darzynkiewicz et al 2014 ) and induction of apoptosis (Refaat et al 2015 ). It has been taken into use against Methicillin-resistant Staphylococcus aureus (MRSA) infections (Stermitz et al 2000 ).…”